Doxorubicin Stability-indicating Method and its Main Degradation Products In vitro Toxicity.

Abstract

Human health and the environment are continuously impacted by anthropogenic activities, particularly those involving emerging compounds. As these compounds are newly identified or not yet fully documented in the literature, comprehensive knowledge of their specific toxicities remains limited. Among these, pharmaceutical compounds are of particular concern, as their mechanisms of action and the effects of their pharmaceutical impurities remain insufficiently understood. In this context, this study aimed to evaluate the behavior of the antineoplastic pharmaceutical doxorubicin (DOX) under stress conditions, including acid and base hydrolysis, oxidation, photolysis, and temperature variations. The goal was to identify the major degradation pathways and elucidate the structures of the main degradation products. A stability-indicating HPLC-DAD-MS method was developed and validated for this purpose. Throughout method development, several degradation products were identified, including 7-deoxydehydrodoxorubicinone, formed through acid hydrolysis, and a major thermal degradation product with a mass-to-charge ratio (m/z) of 530. This thermal degradation product was also detected in analyses of expired pharmaceutical formulations. Furthermore, the in vitro toxicity assessment of samples containing degradation products from thermal decomposition revealed cytotoxic effects on mononuclear cells. These findings underscore the importance of not only understanding the degradation pathways of pharmaceutical compounds but also evaluating the potential environmental and human health impacts of these degradation products.